In response to this NCI solicitation, Rockland proposes to develop a novel method to assay the pharmacodynamic (PO) properties of anti-cancer small molecules targeting the PI3K/mTOR/AKT signaling pathway by quantitatively and independently measuring the phosphorylation of each unique AKT isoform in a validated multiplex immunoassay. The developed PD-AKT-ELISA would offer improved personalized medicine, whereby physicians would prescribe a dosage according to the pharmacodynamic effects of anti-cancer drugs on individual patients, or modify therapeutic modality based on the feedback for each individual Akt isoform. We plan to use a number of anti-cancer drug candidates in this study and to produce reagents, including monoclonal antibodies to measure inactive and active AKT isoforms suitable for a fit-for-use theranostic quantitative biomarker ELISA for AKT. The PO-AKT-ELISA and associated reagents will be produced, optimized, validated and applied to both solid tumor and soft tumor xenograft tissue. The reagents and validated assay will measure all AKT isoforms in its active and inactive states to fill a need not currently addressed by products or technologies commercially available to researchers and clinicians. The end result will be a multiplex assay that can be used by most existing immunoassay readers in both a research and high through-put clinical environment. Assay results will allow physicians to more precisely target dosages or types of drugs that modulate the PI3K/mTOR/AKT pathway, and will ultimately lower treatment costs by more accurately prescribing effective doses of anticancer drugs and potentially shortening the duration of drug treatment regimens.